Ontogeny of hemocyanin function in the dungeness crab cancer magister: hemolymph modulation of hemocyanin oxygen-binding
The in vivo oxygen-binding characteristics of Cancer magister whole hemolymph were compared across developmental stages with those of purified hemocyanin. When the 25S hemocyanins from first-instar juvenile and adult C. magister were dialyzed against first-instar juvenile saline, the P50 values at pH 7.8 differed by 54 %: 2.16 kPa for the adult and 4.68 kPa for the first-instar juvenile. Since both purified proteins were examined under identical conditions, this represents an intrinsic stage-specific difference in hemocyanin O2-affinity. When the two types of hemocyanin were dialyzed against their respective stage-specific salines, the oxygen affinities differed by only 28 %: 3.39 kPa for the adult and still 4.68 kPa for the first-instar juvenile. Thus, the intrinsic difference in hemocyanin O2-affinity was reduced by the stage-specific differences in hemolymph ion concentrations. Even more significant is the fact that the whole-hemolymph P50 values of the juvenile and adult were indistinguishable at in vivo pH and divalent cation levels specific for each stage. Thus, despite significant differences in the intrinsic oxygen affinity of the purified 25S hemocyanin during development, the whole-hemolymph oxygen-binding properties are conserved. In the juvenile crab, it appears that the low-affinity hemocyanin serves to modulate the effects of a weak renal regulation of [Mg2+]. As ion regulation is enhanced during development and divalent cation levels decrease, the crab synthesizes higher-affinity hemocyanin.
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